BIOC14H3 Lecture Notes - Lecture 6: Predictive Validity, Visual Acuity, Lithium Chloride
26 May 2018
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Lecture 6
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Lecture 6 Objectives
• Genome editing using CRISPR/Cas9
• Genetic dissection of neuronal circuits
• RNA in situ hybridization
• Genetic access to a specific neuron population
• Using promoters/enhancers
• Using BACs
• Knock-in approach
• Recombinase-mediated approach
• Virus mediated approach
• CRISPR/Cas9
• Phenotypic analysis of animal behaviors
• General health
• Social behaviors
• Learning and memory
• Spatial navigation
• Aversive learning
• Emotional behaviors
• Fear/anxiety, depression, schizophrenia
related
• Reward seeking behaviors
• Motor functions
• Sensory functions
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Genome editing using CRISPR/CAS9
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• Naturally occurring system in prokaryotes
that defends against viral genetic material
• CRISPR – Clustered Regularly Interspaced
Short Palindromic Repeats à stretch of
DNA that contains short regions à
transcription of these regions makes RNA
that serves as guides (short-guide RNAs or
sgRNAs) to recognize viral DNA segments
• CAS9 – CRISPR associated enzyme 9 à An
endonuclease that binds to sgRNA and
catalyses double-stranded break in foreign
DNA and disables it
• sgRNA can be engineered to recognize any
gene (or DNA sequence) and cause a double-
stranded break
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Document Summary
Social behaviors: general health, learning and memory. Spatial navigation: aversive learning, emotional behaviors, fear/anxiety, depression, schizophrenia related, reward seeking behaviors, motor functions. Genome editing using crispr/cas9: naturally occurring system in prokaryotes that defends against viral genetic material, crispr clustered regularly interspaced. Dna and disables it sgrna can be engineered to recognize any gene (or dna sequence) and cause a double- stranded break. Genome editing using crispr/cas9: cellular systems try to repair double-stranded breaks can be exploited to insert another piece of dna. Rna hybrid: cas9 bound to sgrna cuts the target dna on both strands double-stranded break. Genome editing using crispr/cas9: cells try to repair the double- stranded break using non- homologous end joining (nhej, nhej repair is imperfect and creates indels insertion or deletion of dna bases (2-30 bp) Likely to cause frameshift mutation premature stop codon. Loss of gene function: can be used to target several genes simultaneously. See: pathways of the brain resemble woven sheet of fabric.
